I. General Introduction
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Developing a Genetic Manipulation System for the Antarctic Archaeon, Halorubrum Lacusprofundi: Investigating Acetamidase Gene Function
www.nature.com/scientificreports OPEN Developing a genetic manipulation system for the Antarctic archaeon, Halorubrum lacusprofundi: Received: 27 May 2016 Accepted: 16 September 2016 investigating acetamidase gene Published: 06 October 2016 function Y. Liao1, T. J. Williams1, J. C. Walsh2,3, M. Ji1, A. Poljak4, P. M. G. Curmi2, I. G. Duggin3 & R. Cavicchioli1 No systems have been reported for genetic manipulation of cold-adapted Archaea. Halorubrum lacusprofundi is an important member of Deep Lake, Antarctica (~10% of the population), and is amendable to laboratory cultivation. Here we report the development of a shuttle-vector and targeted gene-knockout system for this species. To investigate the function of acetamidase/formamidase genes, a class of genes not experimentally studied in Archaea, the acetamidase gene, amd3, was disrupted. The wild-type grew on acetamide as a sole source of carbon and nitrogen, but the mutant did not. Acetamidase/formamidase genes were found to form three distinct clades within a broad distribution of Archaea and Bacteria. Genes were present within lineages characterized by aerobic growth in low nutrient environments (e.g. haloarchaea, Starkeya) but absent from lineages containing anaerobes or facultative anaerobes (e.g. methanogens, Epsilonproteobacteria) or parasites of animals and plants (e.g. Chlamydiae). While acetamide is not a well characterized natural substrate, the build-up of plastic pollutants in the environment provides a potential source of introduced acetamide. In view of the extent and pattern of distribution of acetamidase/formamidase sequences within Archaea and Bacteria, we speculate that acetamide from plastics may promote the selection of amd/fmd genes in an increasing number of environmental microorganisms. -
Polyamine Profiles of Some Members of the Alpha Subclass of the Class Proteobacteria: Polyamine Analysis of Twenty Recently Described Genera
Microbiol. Cult. Coll. June 2003. p. 13 ─ 21 Vol. 19, No. 1 Polyamine Profiles of Some Members of the Alpha Subclass of the Class Proteobacteria: Polyamine Analysis of Twenty Recently Described Genera Koei Hamana1)*,Azusa Sakamoto1),Satomi Tachiyanagi1), Eri Terauchi1)and Mariko Takeuchi2) 1)Department of Laboratory Sciences, School of Health Sciences, Faculty of Medicine, Gunma University, 39 ─ 15 Showa-machi 3 ─ chome, Maebashi, Gunma 371 ─ 8514, Japan 2)Institute for Fermentation, Osaka, 17 ─ 85, Juso-honmachi 2 ─ chome, Yodogawa-ku, Osaka, 532 ─ 8686, Japan Cellular polyamines of 41 newly validated or reclassified alpha proteobacteria belonging to 20 genera were analyzed by HPLC. Acetic acid bacteria belonging to the new genus Asaia and the genera Gluconobacter, Gluconacetobacter, Acetobacter and Acidomonas of the alpha ─ 1 sub- group ubiquitously contained spermidine as the major polyamine. Aerobic bacteriochlorophyll a ─ containing Acidisphaera, Craurococcus and Paracraurococcus(alpha ─ 1)and Roseibium (alpha-2)contained spermidine and lacked homospermidine. New Rhizobium species, including some species transferred from the genera Agrobacterium and Allorhizobium, and new Sinorhizobium and Mesorhizobium species of the alpha ─ 2 subgroup contained homospermidine as a major polyamine. Homospermidine was the major polyamine in the genera Oligotropha, Carbophilus, Zavarzinia, Blastobacter, Starkeya and Rhodoblastus of the alpha ─ 2 subgroup. Rhodobaca bogoriensis of the alpha ─ 3 subgroup contained spermidine. Within the alpha ─ 4 sub- group, the genus Sphingomonas has been divided into four clusters, and species of the emended Sphingomonas(cluster I)contained homospermidine whereas those of the three newly described genera Sphingobium, Novosphingobium and Sphingopyxis(corresponding to clusters II, III and IV of the former Sphingomonas)ubiquitously contained spermidine. -
Phosphorus Recovery from Sewage Sludge Using Acidithiobacilli
International Journal of Environmental Research and Public Health Article Phosphorus Recovery from Sewage Sludge Using Acidithiobacilli Surendra K. Pradhan 1,* , Helvi Heinonen-Tanski 1, Anna-Maria Veijalainen 1, Sirpa Peräniemi 2 and Eila Torvinen 1 1 Department of Environmental and Biological Sciences, University of Eastern Finland, FI-70211 Kuopio, Finland; helvi.heinonentanski@uef.fi (H.H.-T.); anna-maria.veijalainen@uef.fi (A.-M.V.); eila.torvinen@uef.fi (E.T.) 2 Department of Pharmacy, University of Eastern Finland, FI-70211 Kuopio, Finland; sirpa.peraniemi@uef.fi * Correspondence: surendra.pradhan@uef.fi Abstract: Sewage sludge contains a significant amount of phosphorus (P), which could be recycled to address the global demand for this non-renewable, important plant nutrient. The P in sludge can be solubilized and recovered so that it can be recycled when needed. This study investigated the P solubilization from sewage sludge using Acidithiobacillus thiooxidans and Acidithiobacillus ferroox- idans. The experiment was conducted by mixing 10 mL of sewage sludge with 90 mL of different water/liquid medium/inoculum and incubated at 30 ◦C. The experiment was conducted in three semi-continuous phases by replacing 10% of the mixed incubated medium with fresh sewage sludge. In addition, 10 g/L elemental sulfur (S) was supplemented into the medium in the third phase. The pH of the A. thiooxidans and A. ferrooxidans treated sludge solutions was between 2.2 and 6.3 until day 42. In phase 3, after supplementing with S, the pH of A. thiooxidans treated sludge was reduced to 0.9, Citation: Pradhan, S.K.; which solubilized and extracted 92% of P. -
Rubisco POSTER 2016 MACUB
Direct PCR Detection, Cloning, and Characterization of Bacterial RubisCO Genes from New Jersey Soils Stephanie Zapata*, Anna Gonzalez, Margarita Kulko, Ryan Kim, Theranda Jashari, Aidan Holwerda, Tina Choe, and Luis Jimenez Department of Biology and Horticulture, Bergen Community College, Paramus, New Jersey, USA Abstract Materials and Methods Ribulose-1,5-bisphosphate carboxylase/oxygenase, commonly known by PCR detection of bacterial RubisCO genes in soil the abbreviation RubisCO, is an enzyme involved in the first major step of Cloning libraries carbon fixation, a process by which atmospheric carbon dioxide is The DNA fragments from the PCR amplification of RubisCO converted by bacteria to energy-rich molecules such as glucose. genes were cloned using plasmid pCR®4-TOPO (Life Microbial DNA was extracted from temperate soils using the Zymo Technologies, Thermo Fisher Scientific, Grand Island, NY) Microbe DNA MiniPrep protocol. RubisCo gene sequences were according to the manufacturer’s instructions. Transformations amplified by PCR using degenerate primers cbbLG1F and cbbLG1R. were performed using Mix and Go Competent E. coli strains DNA fragments of approximately 800 base pair were detected in all positive soil samples. Clone libraries were constructed with the amplified (Zymo Research, Irvine, CA). White colonies grown on Luria ç800 bp DNA fragments by ligating the detected fragments with vector pCR®4- Bertani (LB) Agar with ampicillin (50 ug/ml) were transferred to LB TOPO. Transformations were performed using competent Mix and Go broth containing ampicillin (50 ug/ml). Samples were incubated Escherichia coli cells. Plasmids were isolated from each clone using the overnight at 37°C. Zyppy Plasmid Miniprep and inserts were screened by PCR using M13 Plasmids were isolated from each clone using the Zyppy Plasmid DNA primers. -
A Survey of Carbon Fixation Pathways Through a Quantitative Lens
Journal of Experimental Botany, Vol. 63, No. 6, pp. 2325–2342, 2012 doi:10.1093/jxb/err417 Advance Access publication 26 December, 2011 REVIEW PAPER A survey of carbon fixation pathways through a quantitative lens Arren Bar-Even, Elad Noor and Ron Milo* Department of Plant Sciences, The Weizmann Institute of Science, Rehovot 76100, Israel * To whom correspondence should be addressed. E-mail: [email protected] Received 15 August 2011; Revised 4 November 2011; Accepted 8 November 2011 Downloaded from Abstract While the reductive pentose phosphate cycle is responsible for the fixation of most of the carbon in the biosphere, it http://jxb.oxfordjournals.org/ has several natural substitutes. In fact, due to the characterization of three new carbon fixation pathways in the last decade, the diversity of known metabolic solutions for autotrophic growth has doubled. In this review, the different pathways are analysed and compared according to various criteria, trying to connect each of the different metabolic alternatives to suitable environments or metabolic goals. The different roles of carbon fixation are discussed; in addition to sustaining autotrophic growth it can also be used for energy conservation and as an electron sink for the recycling of reduced electron carriers. Our main focus in this review is on thermodynamic and kinetic aspects, including thermodynamically challenging reactions, the ATP requirement of each pathway, energetic constraints on carbon fixation, and factors that are expected to limit the rate of the pathways. Finally, possible metabolic structures at Weizmann Institute of Science on July 3, 2016 of yet unknown carbon fixation pathways are suggested and discussed. -
Comparative Genome Analysis of Acidithiobacillus Ferrooxidans, A
Hydrometallurgy 94 (2008) 180–184 Contents lists available at ScienceDirect Hydrometallurgy journal homepage: www.elsevier.com/locate/hydromet Comparative genome analysis of Acidithiobacillus ferrooxidans, A. thiooxidans and A. caldus: Insights into their metabolism and ecophysiology Jorge Valdés 1, Inti Pedroso, Raquel Quatrini, David S. Holmes ⁎ Center for Bioinformatics and Genome Biology, Life Science Foundation, MIFAB and Andrés Bello University, Santiago, Chile ARTICLE INFO ABSTRACT Available online 2 July 2008 Draft genome sequences of Acidthiobacillus thiooxidans and A. caldus have been annotated and compared to the previously annotated genome of A. ferrooxidans. This has allowed the prediction of metabolic and Keywords: regulatory models for each species and has provided a unique opportunity to undertake comparative Acidithiobacillus genomic studies of this group of related bioleaching bacteria. In this paper, the presence or absence of Bioinformatics predicted genes for eleven metabolic processes, electron transfer pathways and other phenotypic Comparative genomics fi Metabolic integration characteristics are reported for the three acidithiobacilli: CO2 xation, the TCA cycle, sulfur oxidation, sulfur reduction, iron oxidation, iron assimilation, quorum sensing via the acyl homoserine lactone mechanism, hydrogen oxidation, flagella formation, Che signaling (chemotaxis) and nitrogen fixation. Predicted transcriptional and metabolic interplay between pathways pinpoints possible coordinated responses to environmental signals such as energy source, oxygen and nutrient limitations. The predicted pathway for nitrogen fixation in A. ferrooxidans will be described as an example of such an integrated response. Several responses appear to be especially characteristic of autotrophic microorganisms and may have direct implications for metabolic processes of critical relevance to the understanding of how these microorganisms survive and proliferate in extreme environments, including industrial bioleaching operations. -
1 Characterization of Sulfur Metabolizing Microbes in a Cold Saline Microbial Mat of the Canadian High Arctic Raven Comery Mast
Characterization of sulfur metabolizing microbes in a cold saline microbial mat of the Canadian High Arctic Raven Comery Master of Science Department of Natural Resource Sciences Unit: Microbiology McGill University, Montreal July 2015 A thesis submitted to McGill University in partial fulfillment of the requirements of the degree of Master in Science © Raven Comery 2015 1 Abstract/Résumé The Gypsum Hill (GH) spring system is located on Axel Heiberg Island of the High Arctic, perennially discharging cold hypersaline water rich in sulfur compounds. Microbial mats are found adjacent to channels of the GH springs. This thesis is the first detailed analysis of the Gypsum Hill spring microbial mats and their microbial diversity. Physicochemical analyses of the water saturating the GH spring microbial mat show that in summer it is cold (9°C), hypersaline (5.6%), and contains sulfide (0-10 ppm) and thiosulfate (>50 ppm). Pyrosequencing analyses were carried out on both 16S rRNA transcripts (i.e. cDNA) and genes (i.e. DNA) to investigate the mat’s community composition, diversity, and putatively active members. In order to investigate the sulfate reducing community in detail, the sulfite reductase gene and its transcript were also sequenced. Finally, enrichment cultures for sulfate/sulfur reducing bacteria were set up and monitored for sulfide production at cold temperatures. Overall, sulfur metabolism was found to be an important component of the GH microbial mat system, particularly the active fraction, as 49% of DNA and 77% of cDNA from bacterial 16S rRNA gene libraries were classified as taxa capable of the reduction or oxidation of sulfur compounds. -
Acidithiobacillus Ferrooxidans for Biotechnological Applications
Engineering and Characterization of Acidithiobacillus ferrooxidans for Biotechnological Applications Xiaozheng Li Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2016 © 2015 Xiaozheng Li All rights reserved ABSTRACT Engineering and Characterization of Acidithiobacillus ferrooxidans for Biotechnology Applications Xiaozheng Li Acidithiobacillus ferrooxidans is a gram-negative bacterium that is able to extract energy from oxidation of Fe 2+ and reduced sulfur compounds and fix carbon dioxide from atmosphere. The facts that A. ferrooxidans thrives in acidic pH (~2), fixes carbon dioxide from the atmosphere and oxidizes Fe 2+ for energy make it a good candidate in many industrial applications such as electrofuels and biomining. Electrofuels is a new type of bioprocess, which aims to store electrical energy, such as solar power, in the form of chemical bonds in the liquid fuels. Unlike traditional biofuels made from agricultural feedstocks, electrofuels bypass the inefficient photosynthesis process and thus have potentially higher photon-to-fuel efficiency than traditional biofuels. This thesis covers the development of a novel bioprocess involving A. ferrooxidans to make electrofuels, i.e. isobutyric acid and heptadecane. There are four major steps: characterization of wild-type cells, engineering of medium for improved electrochemical performance, genetic modification of A. ferrooxidans and optimization of operating conditions to enhance biofuel production. Each is addressed in one of the chapters in this thesis. In addition, applications of A. ferrooxidans in biomining processes will be briefly discussed. An economic analysis of various applications including electrofuels and biomining is also presented. Wild-type A. -
Microbial Ecology of Biofiltration Units Used for the Desulfurization of Biogas
chemengineering Review Microbial Ecology of Biofiltration Units Used for the Desulfurization of Biogas Sylvie Le Borgne 1,* and Guillermo Baquerizo 2,3 1 Departamento de Procesos y Tecnología, Universidad Autónoma Metropolitana- Unidad Cuajimalpa, 05348 Ciudad de México, Mexico 2 Irstea, UR REVERSAAL, F-69626 Villeurbanne CEDEX, France 3 Departamento de Ingeniería Química, Alimentos y Ambiental, Universidad de las Américas Puebla, 72810 San Andrés Cholula, Puebla, Mexico * Correspondence: [email protected]; Tel.: +52–555–146–500 (ext. 3877) Received: 1 May 2019; Accepted: 28 June 2019; Published: 7 August 2019 Abstract: Bacterial communities’ composition, activity and robustness determines the effectiveness of biofiltration units for the desulfurization of biogas. It is therefore important to get a better understanding of the bacterial communities that coexist in biofiltration units under different operational conditions for the removal of H2S, the main reduced sulfur compound to eliminate in biogas. This review presents the main characteristics of sulfur-oxidizing chemotrophic bacteria that are the base of the biological transformation of H2S to innocuous products in biofilters. A survey of the existing biofiltration technologies in relation to H2S elimination is then presented followed by a review of the microbial ecology studies performed to date on biotrickling filter units for the treatment of H2S in biogas under aerobic and anoxic conditions. Keywords: biogas; desulfurization; hydrogen sulfide; sulfur-oxidizing bacteria; biofiltration; biotrickling filters; anoxic biofiltration; autotrophic denitrification; microbial ecology; molecular techniques 1. Introduction Biogas is a promising renewable energy source that could contribute to regional economic growth due to its indigenous local-based production together with reduced greenhouse gas emissions [1]. -
Molecular Systematics of the Genus Acidithiobacillus: Insights Into the Phylogenetic Structure and Diversification of the Taxon
ORIGINAL RESEARCH published: 19 January 2017 doi: 10.3389/fmicb.2017.00030 Molecular Systematics of the Genus Acidithiobacillus: Insights into the Phylogenetic Structure and Diversification of the Taxon Harold Nuñez 1 †, Ana Moya-Beltrán 1, 2 †, Paulo C. Covarrubias 1, Francisco Issotta 1, Juan Pablo Cárdenas 3, Mónica González 1, Joaquín Atavales 1, Lillian G. Acuña 1, D. Barrie Johnson 4* and Raquel Quatrini 1* 1 Microbial Ecophysiology Laboratory, Fundación Ciencia & Vida, Santiago, Chile, 2 Faculty of Biological Sciences, Andres Bello University, Santiago, Chile, 3 uBiome, Inc., San Francisco, CA, USA, 4 College of Natural Sciences, Bangor University, Bangor, UK The acidithiobacilli are sulfur-oxidizing acidophilic bacteria that thrive in both natural and anthropogenic low pH environments. They contribute to processes that lead to the generation of acid rock drainage in several different geoclimatic contexts, and their Edited by: Jesse G. Dillon, properties have long been harnessed for the biotechnological processing of minerals. California State University, Long Presently, the genus is composed of seven validated species, described between 1922 Beach, USA and 2015: Acidithiobacillus thiooxidans, A. ferrooxidans, A. albertensis, A. caldus, A. Reviewed by: Daniel Seth Jones, ferrivorans, A. ferridurans, and A. ferriphilus. However, a large number of Acidithiobacillus University of Minnesota, USA strains and sequence clones have been obtained from a variety of ecological niches over Stephanus Nicolaas Venter, the years, and many isolates are thought to vary in phenotypic properties and cognate University of Pretoria, South Africa genetic traits. Moreover, many isolates remain unclassified and several conflicting specific *Correspondence: D. Barrie Johnson assignments muddle the picture from an evolutionary standpoint. -
The Diversity of Cultivable Hydrocarbon-Degrading Bacteria Isolated from Crude Oil Contaminated Soil and Sludge from Arzew Refinery in Algeria
THE DIVERSITY OF CULTIVABLE HYDROCARBON-DEGRADING BACTERIA ISOLATED FROM CRUDE OIL CONTAMINATED SOIL AND SLUDGE FROM ARZEW REFINERY IN ALGERIA Sonia SEKKOUR1*, Abdelkader BEKKI1, Zoulikha BOUCHIBA1, Timothy M. Vogel2, Elisabeth NAVARRO2 Address(es): Ing. Sonia SEKKOUR PhD., 1Université Ahmed Benbella, Faculté des sciences de la nature et de la vie, Département de Biotechnologie, Laboratoire de biotechnologie des rhizobiums et amélioration des plantes, 31000 Oran, Algérie. 2Environmental Microbial Genomics Group, Laboratoire Ampère, Centre National de la Recherche Scientifique, UMR5005, Institut National de la Recherche Agronomique, USC1407, Ecole Centrale de Lyon, Université de Lyon, Ecully, France. *Corresponding author: [email protected] doi: 10.15414/jmbfs.2019.9.1.70-77 ARTICLE INFO ABSTRACT Received 27. 3. 2018 The use of autochtonious bacterial strains is a valuable bioremediation strategy for cleaning the environment from hydrocarbon Revised 19. 2. 2019 pollutants. The isolation, selection and identification of hydrocarbon-degrading bacteria is therefore crucial for obtaining the most Accepted 14. 3. 2019 promising strains for decontaminate a specific site. In this study, two different media, a minimal medium supplemented with petroleum Published 1. 8. 2019 and with oil refinery sludge as sole carbon source, were used for the isolation of native hydrocarbon-degrading bacterial strains from crude oil contaminated soils and oil refinery sludges which allowed isolation of fifty-eight strains.The evalution of diversity of twenty- two bacterials isolates reveled a dominance of the phylum Proteobacteria (20/22 strains), with a unique class of Alphaproteobacteria, Regular article the two remaining strains belong to the phylum Actinobacteria. Partial 16S rRNA gene sequencing performed on isolates showed high level of identity with known sequences. -
Thermithiobacillus Tepidarius DSM 3134T, a Moderately Thermophilic, Obligately Chemolithoautotrophic Member of the Acidithiobacillia
Boden et al. Standards in Genomic Sciences (2016) 11:74 DOI 10.1186/s40793-016-0188-0 SHORT GENOME REPORT Open Access Permanent draft genome of Thermithiobacillus tepidarius DSM 3134T, a moderately thermophilic, obligately chemolithoautotrophic member of the Acidithiobacillia Rich Boden1,2* , Lee P. Hutt1,2, Marcel Huntemann3, Alicia Clum3, Manoj Pillay3, Krishnaveni Palaniappan3, Neha Varghese3, Natalia Mikhailova3, Dimitrios Stamatis3, Tatiparthi Reddy3, Chew Yee Ngan3, Chris Daum3, Nicole Shapiro3, Victor Markowitz3, Natalia Ivanova3, Tanja Woyke3 and Nikos Kyrpides3 Abstract Thermithiobacillus tepidarius DSM 3134T was originally isolated (1983) from the waters of a sulfidic spring entering the Roman Baths (Temple of Sulis-Minerva) at Bath, United Kingdom and is an obligate chemolithoautotroph growing at the expense of reduced sulfur species. This strain has a genome size of 2,958,498 bp. Here we report the genome sequence, annotation and characteristics. The genome comprises 2,902 protein coding and 66 RNA coding genes. Genes responsible for the transaldolase variant of the Calvin-Benson-Bassham cycle were identified along with a biosynthetic horseshoe in lieu of Krebs’ cycle sensu stricto. Terminal oxidases were identified, viz. cytochrome c oxidase (cbb3, EC 1.9.3.1) and ubiquinol oxidase (bd, EC 1.10.3.10). Metalloresistance genes involved in pathways of arsenic and cadmium resistance were found. Evidence of horizontal gene transfer accounting for 5.9 % of the protein-coding genes was found, including transfer from Thiobacillus spp. and Methylococcus capsulatus Bath, isolated from the same spring. A sox gene cluster was found, similar in structure to those from other Acidithiobacillia – by comparison with Thiobacillus thioparus and Paracoccus denitrificans, an additional gene between soxA and soxB was found, annotated as a DUF302-family protein of unknown function.